• 제목/요약/키워드: Amplified Fragment Length Polymorphism

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Distinction between Cold-sensitive and -tolerant Jute by DNA Polymorphisms

  • Hossain, Mohammad Belayat;Awal, Aleya;Rahman, Mohammad Aminur;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • v.36 no.5
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    • pp.427-432
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    • 2003
  • Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

Analysis of genetic relationships of Colletotrichum spp. isolated from sweet persimon with AFLP (AFLP를 이용한 단감나무 탄저병 병원균 Colletotrichum spp.의 유연관계 분석)

  • Kim, Hee-Jong;Jeong, Bong-Gu;Lee, Youn-Su
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.9-14
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    • 2001
  • Colletotrichum species are important fungal pathogens that cause great damages on various host plant species worldwide. In Korea, Colletotrichum species cause massive economic losses on apple, peach, grape, and especially, sweet persimon productions. In the past, identification of the pathogen and the studies on the genetic relationships among the pathogenic isolates were mainly based on morphology, cultural characteristics, and the difference in pathogenicity. However, in recent years, these traditional methods have been replaced with molecular methods including AFLP. AFLP method with the merits of both RAPD and RFLP has been widely used for the genetic relationship studies of various organisms. Therefore, in this study, AFLP method was employed for the studies of genetic relationships among the different isolates of Colletotrichum species collected from various parts of sothern Korea. As a result, two specific band pattern groups were observed among different isolates of Colletotrichum species.

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A phytogeographical study of Sasa borealis populations based on AFLP analysis (AFLP 마커를 이용한 조릿대 개체군의 식물지리학적 연구)

  • Kim, Il Ryong;Yu, Dasom;Choi, Hong-Keun
    • Korean Journal of Plant Taxonomy
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    • v.45 no.1
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    • pp.29-35
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    • 2015
  • Sasa borealis (Hack.) Makino & Shibata is widely distributed in South Korea. With amplified fragment length polymorphism (AFLP) markers, we analyzed the genetic diversity of S. borealis to predict and measure the phytogeographical factors of these populations. Relatively high levels of genetic diversity (PPL = 37.2%, h = 0.143, I = 0.205) and genetic differentiation ($G_{ST}$ = 0.324, ${\theta}^B$ = 0.395) were confirmed in populations of S. borealis. Moreover, an analysis of molecular variance (AMOVA) showed that the rate of differentiation among the populations was 47.7%. The results showed that genetic diversity is inversely proportional to the latitude of the S. borealis populations, indicating that the distribution of S. borealis may have extended from lower to higher latitudes. This method of investigating the correlation between genetic diversity and latitude presents critical information for estimating changes in distributions and plant conservation due to climate change.

A unique genetic lineage at the southern coast of China in the agar-producing Gracilaria vermiculophylla (Gracilariales, Florideophyceae)

  • Hu, Zi-Min;Liu, Ruo-Yu;Zhang, Jie;Duan, De-Lin;Wang, Gao-Ge;Li, Wen-Hong
    • ALGAE
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    • v.33 no.3
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    • pp.269-278
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    • 2018
  • Ocean warming can have significant negative impacts on population genetic diversity, local endemism and geographical distribution of a wide range of marine organisms. Thus, the identification of conservation units with high risk of extinction becomes an imperative task to assess, monitor, and manage marine biodiversity for policy-makers. Here, we surveyed population structure and genetic variation of the red seaweed Gracilaria vermiculophylla along the coast of China using genome-based amplified fragment length polymorphism (AFLP) scanning. Regardless of analysis methods used, AFLP consistently revealed a south to north genetic isolation. Populations at the southern coast of China showed unique genetic variation and much greater allelic richness, heterozygosity, and average genetic diversity than the northern. In particular, we identified a geographical barrier that may hinder genetic exchange between the two lineages. Consequently, the characterized genetic lineage at the southern coast of China likely resulted from the interplay of post-glacial persistence of ancestral diversity, geographical isolation and local adaptation. In particular, the southern populations are indispensable components to explore evolutionary genetics and historical biogeography of G. vermiculophylla in the northwestern Pacific, and the unique diversity also has important conservation value in terms of projected climate warming.

Application of Methodology for Microbial Community Analysis to Gas-Phase Biofilters (폐가스 처리용 바이오필터에 미생물 군집 분석 기법의 적용)

  • Lee, Eun-Hee;Park, Hyunjung;Jo, Yun-Seong;Ryu, Hee Wook;Cho, Kyung-Suk
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.147-156
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    • 2010
  • There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

Identification of Luteovirus Nucleotide Sequences in Mild Yellow-Edge Diseased Strawberry Plants

  • Shaban Montasser, Magdy;Al-Awadhi, Husain;Hadidi, Ahmed
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.1-5
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    • 2002
  • The availability of nucleotide sequences of the coat protein gene of Potato leafroll virus (PLRV) permitted the construction of DNA primers that were utilized for cDNA synthesis. Polymerase chain reaction (PCR) products of a 487 bp. and approximately 500 bp DNA fragments were amplified from nucleic acid extracts of PLRV-infected tissue and strawberry mild yellow-edge (SMYE) diseased strawberry tissue, respectively. The amplified DNA fragments were further differentiated by hybridization analysis with a CDNA probe for the coat protein gene of PLRV and restriction fragment length polymorphism (RFLP) analysis. These results suggest that a luteovirus is associated with the SMYE disease.

Comparative AFLP Profiles among Strains of Korean Races of Xanthomonas oryzae pv. oryzae.

  • Kang, Mi-Hyung;Lee, Du-Ku;Noh, Tae-Hwan;Shim, Hyeong-Kwon;Na, Seung-Yong;Kim, Jae-Duk
    • Plant Resources
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    • v.7 no.1
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    • pp.65-68
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    • 2004
  • We used an amplified fragment length polymorphism (AFLP) analysis, a novel PCR-based technique, to differentiate Xanthomonas oryzae pv. oryzae (Xoo) of Korean races. The 6 strains of Xoo K1, K2, K3 races were tested with 81 AFLP primer combinations to identify the best selective primers. The primer combinations were selected according to their reproducibility, number of polymorphic bands and polymorphism detected among Xoo strains. 18 strains of Xoo K1, K2 and K3 races were analyzed with the selected combinations of primer set. Some primer combinations (Eco R I +1 / Mse I+1) could differentiate Xoo of Korean races that were not distinguished by other fingerprinting analysis. Thus AFLP fingerprinting permitted very fine discrimination among different races.

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Genetic Diversity of Korean Cyanobacteria determined by DNA polymorphisms within the Phycocyanin Locus (Phycocyanin locus내의 DNA Polymorphism에 의한 한국산 Cyanobacteria의 유전적 다양성)

  • 박진숙;권주리;유순애
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.249-253
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    • 2000
  • The genetic diversity among Korean cyanobacteria was assessed by restriction fragment length polymorphism(RFLP) analysis of PCR products from the phycocyanin locus. Strains of all the genera tested were successfully amplified, and the size of amplified fragments was approximately 700bp. The restriction patterns generated by AluI, MspI, and HaeIII were conserved for strains within each of genera studied and were specific to the genus level. Intrageneric delineation of strains was revealed by the enzyme, CfoI for members of genera Anabeana and Synechocystis. Phenogram derived from the different RFLP patterns revealed a coherent cluster among Anabeana, Chlorogloea, and Synechocystis strains. PC-RFLP methods provided useful tools for classification of cyanobacteria.

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Identification of Pork Contamination in Meatballs of Indonesia Local Market Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis

  • Erwanto, Yuny;Abidin, Mohammad Zainal;Muslim, Eko Yasin Prasetyo;Sugiyono, Sugiyono;Rohman, Abdul
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.10
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    • pp.1487-1492
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    • 2014
  • This research applied and evaluated a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using cytochrome b gene to detect pork contamination in meatballs from local markets in Surabaya and Yogyakarta regions, Indonesia. To confirm the effectiveness and specificity of this fragment, thirty nine DNA samples from different meatball shops were isolated and amplified, and then the PCR amplicon was digested by BseDI restriction enzyme to detect the presence of pork in meatballs. BseDI restriction enzyme was able to cleave porcine cytochrome b gene into two fragments (131 bp and 228 bp). Testing the meatballs from the local market showed that nine of twenty meatball shops in Yogyakarta region were detected to have pork contamination, but there was no pork contamination in meatball shops in Surabaya region. In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity. Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked. The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

An Overview of Different Techniques on the Microbial Community Structure, and Functional Diversity of Plant Growth Promoting Bacteria

  • Kim, Kiyoon;Islam, Rashedul;Benson, Abitha;Joe, Manoharan Melvin;Denver, Walitang;Chanratan, Mak;Chatterjee, Poulami;Kang, Yeongyeong;Sa, Tongmin
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.2
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    • pp.144-156
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    • 2016
  • Soil is a dynamic biological system, in which it is difficult to determine the composition of microbial communities. Knowledge of microbial diversity and function in soils are limited because of the taxonomic and methodological limitations associated with studying the organisms. In this review, approaches to measure microbial diversity in soil were discussed. Research on soil microbes can be categorized as structural diversity, functional diversity and genetic diversity studies, and these include cultivation based and cultivation independent methods. Cultivation independent technique to evaluate soil structural diversity include different techniques such as Phospholipid Fatty Acids (PLFA) and Fatty Acid Methyl Ester (FAME) analysis. Carbon source utilization pattern of soil microorganisms by Community Level Physiological Profiling (CLPP), catabolic responses by Substrate Induced Respiration technique (SIR) and soil microbial enzyme activities are discussed. Genetic diversity of soil microorganisms using molecular techniques such as 16S rDNA analysis Denaturing Gradient Gel Electrophoresis (DGGE) / Temperature Gradient Gel Electrophoresis (TGGE), Terminal Restriction Fragment Length Polymorphism (T-RFLP), Single Strand Conformation Polymorphism (SSCP), Restriction Fragment Length Polymorphism (RFLP) / Amplified Ribosomal DNA Restriction Analysis (ARDRA) and Ribosomal Intergenic Spacer Analysis (RISA) are also discussed. The chapter ends with a final conclusion on the advantages and disadvantages of different techniques and advances in molecular techniques to study the soil microbial diversity.